Biomedical subjects

J B Yoon

Publications and source records attributed to J B Yoon.

At least 19 recordsLinked to original sources

PubMed2001

Non-apoptotic signaling pathways activated by soluble Fas ligand in serum-starved human fibroblasts. Mitogen-activated protein kinases and NF-kappaB-dependent gene expression.

Many Fas-expressing cells do not undergo cell death upon Fas stimulation. In the normal human diploid cell line GM6112, the addition of soluble Fas ligand (sFasL) leads to morphological signs of cell death in less than 1% of cells. Treatment of serum-starved GM6112 fibroblasts with sFasL resulted in a rapid and transient phosphorylation of ERK1/2 without a significant increase in JNK and p38 activities. Unless co-treated with the protein synthesis inhibitor anisomycin, sFasL did not show gene-inducing activity in cells maintained in complete medium. However, when cells were serum-starved for 4 days, treatment with sFasL alone induced interleukin-6 gene expression and, less strongly, interleukin-8 gene expression. Sensitization of the gene-inducing activity by serum starvation correlated with NF-kappaB activation by sFasL. Furthermore, we found that the expression of FADD and caspase-8 was significantly reduced in serum-starved cells, whereas the level of cFLIP remained unchanged. Transfection of GM6112 cells with the antisense caspase-8 expression construct sensitized cells toward sFasL-induced NF-kappaB-dependent reporter activation. Our results support the notion that a change in the ratio of cFLIP and caspase-8 may be responsible for turning on the Fas-activated NF-kappaB pathway, which otherwise is supplanted by the death-inducing pathway.

Animals↗

PubMed2001

Co K-edge XAS study on a new cobalt-doped-SiO2 pillared clay.

New Co-doped-SiO2-sol pillared montmorillonite has been synthesized by interlayer hydrolysis and condensation of tetraethylorthosilicate (TEOS) in the presence of Co2+ ion using an organic template. The Co K-edge XANES and EXAFS analyses for the CoO-SiO2-PILC (before and after calcination) and for the references such as CoO and Co(OH)2 were performed in order to obtain electronic and local structural information of cobalt species, which may act as a catalytic active site for the selective reduction of NO(x), hydrodesulfurization, and Fischer-Tropsch reaction. According to the XANES spectra, the divalent cobalt ion is stabilized in an octahedral symmetry. The EXAFS result shows a significant change in local symmetry around cobalt ion upon calcination. The EXAFS fitting result before calcination shows that the cobalt species is in the form of hydroxide, with a small number of (Co-Co) pairs compared to the bulk Co(OH)2. After calcining at 550 degrees C, the first nearest neighbours were fitted to six oxygen atoms with two different distances, and the second and the third neighbours were fitted to two Si and one Co atoms. It is, therefore, reasonable to suggest a structure model, where the cobalt species on the SiO2 sol exists as a nano cluster of Co(OH)2 before calcination but transforms to a nanosized cobalt oxide covalently bound to the surface of SiO2 pillar after calcination.

Journal Article↗

PubMed2001

In situ XAFS study at the Zr K-edge for SiO2/ZrO2 nano-sol.

The structural characterisation of SiO2/ZrO2 nano-sol particles, prepared by mixing SiO2 sol and aqueous solution of ZrOCl2 8H2O, has been carried out by in-situ XAS measurement at the Zr K-edge during condensation reaction. The detailed XANES features at the Zr K-edge of the mixed sol of SiO2/ZrO2 are compared with those of other references such as ZrO2, ZrOCl2 8H2O. BaZrO3, and ZrSiO4, and it becomes obvious that the Zr4+ ions are stabilised in an octahedral symmetry. The EXAFS result also indicates that each Zr atom is coordinated with six oxygen ones as the first nearest neighbour, where two oxygen atoms are from the linkage of (Si-O-Zr) at short distance, and four ones are from water molecules at long distance. As the condensation reaction proceeds, it is found that the number of oxygen atoms due to the formation of (Si-O-Zr) bond at short distance and the second neighbour of silicon atoms increase simultaneously. From the above EXAFS and XANES results, the structural and gelating models could be proposed, which is based on the octahedrally coordinated but distorted zirconium species attaching on the SiO2 sol surface.

Journal Article↗

PubMed2001

Solution structures of the N-terminal domain of histone H4.

Histones, nuclear proteins that interact with DNA to form nucleosomes, are essential for both the regulation of transcription and the packaging of DNA within chromosomes. The N-terminal domain of histone H4 contains four acetylation sites at lysine residues and may play a separate role in chromatin structure from the remainder of the H4 chain. We performed circular dichroism and NMR characterization of both native (H4NTP) and acetylated (Ace-H4NTP) peptides containing N-terminal acetylation domain of histone H4 for various pH environments. Data from CD and NMR suggested that H4NTP exhibited a pH-dependent conformational change, whereas the Ace-H4NTP is insensitive to pH change. However, both peptides showed a defined structural form at acidic pH environments. The solution structure for Ace-H4NTP shows two structurally independent regions comprising residues of Leu10-Gly13 and Arg19-Leu22, demonstrating relatively well-defined turn-type structures. Our results suggest that N-terminal acetylated region of H4 prefers an extended backbone conformation at neutral pH, however, upon acetylation, the regions containing lysine residues induce structural transition, having defined structural form for its optimum function.

Acetyltransferases↗

PubMed2001

The prevalence and types of androgenetic alopecia in Korean men and women.

BACKGROUND: There are racial differences in the prevalence and types of androgenetic alopecia (AGA). There have been several reports on the prevalence and types of AGA in the general population of caucasians, but few studies on Koreans with samples of sufficient numbers have been reported. OBJECTIVES: To obtain a more precise estimate of the prevalence and types of AGA in Korean men and women and to compare the results with those in caucasians. METHODS: The prevalence and types of AGA were analysed in 10,132 Koreans (5531 men and 4601 women) who had visited the Health Examination Centre at Kyung Hee University Hospital for regular health examinations between December 1997 and July 1999. To classify the degree of hair loss for each subject, the Norwood classification was used in men and the Ludwig classification in women. For AGA in men, 'female pattern' was added to the Norwood classification. RESULTS: In Korean men, the prevalence of AGA (Norwood III or above) at all ages was 14.1%. It increased steadily with advancing age, but was lower than that of caucasians: 2.3% in the third decade, 4.0% in the fourth decade, 10.8% in the fifth decade, 24.5% in the sixth decade, 34.3% in the seventh decade and 46.9% over 70 years. Type III vertex involvement was the most common type in the third decade to the seventh decade; over 70 years, type VI was most common. A 'female pattern' was observed in 11.1% of cases. In Korean women, the prevalence of AGA (Ludwig I or above) at all ages was 5.6%. It also increased steadily with advancing age: 0.2% in the third decade, 2.3% in the fourth decade, 3.8% in the fifth decade, 7.4% in the sixth decade, 11.7% in the seventh decade and 24.7% over 70 years. Grade I was the most common type up to the sixth decade; over 60 years, grade I and II were similar in prevalence. Grade III (total baldness) was not observed. A family history of baldness was present in 48.5% of men and 45.2% of women with AGA. CONCLUSIONS: The prevalence of AGA in Korean men and women was lower than that in caucasians, as recorded in the literature. Korean men tend to have more frontal hairline preservation and show a more 'female pattern' of hair thinning than caucasians. Therefore, 'female pattern' should be added to the classification of AGA.

Adult↗

PubMed1998

Cloning and characterization of the 5'-flanking region for the human topoisomerase III gene.

The human DNA topoisomerase III (hTOP3) gene encodes a topoisomerase homologous to the Escherichia coli DNA topoisomerase I subfamily. To understand the mechanisms responsible for regulating hTOP3 expression, we have cloned the 5'-flanking region of the gene coding for the hTOP3 and analyzed its promoter activity. The presence of a single transcription initiation site was suggested by primer extension analysis. The hTOP3 gene promoter is moderately high in GC content and lacks a canonical TATA box, suggesting that hTOP3 promoter has overall similarity to promoters of a number of housekeeping genes. Examination of the promoter sequence indicated the presence of four Sp-1 consensus binding sequences and a putative initiator element surrounding the transcription initiation site. Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the 52-base pair region from -326 to -275 upstream of the transcription initiation site includes a positive cis-acting element(s) for the efficient expression of hTOP3 gene. On the basis of gel mobility shift and supershift assays, we demonstrated that both YY1 and USF1 transcription factors can bind to the 52-base pair region. When HeLa cells were transiently transfected with a mutant construct which had disabled both YY1- and USF1-binding sites, the luciferase activity was greatly reduced, suggesting that these binding elements play a functional role in the basal activation of the hTOP3 promoter. Transfection studies with mutations that selectively impaired YY1 or USF1 binding suggested that both YY1 and USF1 function as activators in the hTOP3 promoter.

Base Sequence↗

PubMed1998

Regional and temporal specialization in the nucleus: a transcriptionally-active nuclear domain rich in PTF, Oct1 and PIKA antigens associates with specific chromosomes early in the cell cycle.

PTF (PSE-binding transcription factor) activates transcription of snRNA and related genes. We investigated its distribution in HeLa nuclei by immunofluorescence, and found it spread throughout the nucleoplasm in small foci. In some cells, PTF is also concentrated in one, or very few, discrete regions (diameter approximately 1.3 micron) that appear during G1 phase and disappear in S phase. Oct1, a transcription factor that interacts with PTF, is also enriched in these domains; RNA polymerase II, TBP and Sp1 are also present. Each domain typically contains 2 or 3 transcription 'factories' where Br-UTP is incorporated into nascent transcripts. Accordingly, we have christened this region the Oct1/PTF/transcription (OPT) domain. It colocalizes with some, but not all, PIKA domains. It is distinct from other nuclear domains, including coiled bodies, gemini bodies, PML bodies and the perinucleolar compartment. A small region on chromosome 6 (band 6p21) containing only approximately 30 Mbp DNA, and chromosomes 6 and 7, associate with the domain significantly more than other chromosomes. The domains may act like nucleoli to bring particular genes on specific chromosomes together to a region where the appropriate transcription and processing factors are concentrated, thereby facilitating the expression of those genes.

Amanitins↗

PubMed1998

The human PTFgamma/SNAP43 gene: structure, chromosomal location, and identification of a VNTR in 5'-UTR.

PTF/SNAPc is a multisubunit complex which specifically recognizes the PSEs of small nuclear RNA genes and activates transcription by RNA polymerase II or III. Here we describe the isolation and characterization of genomic clones encoding the human PTFgamma/SNAP43 gene. The gene spans approximately 29 kilobases, and is composed of 9 exons and 8 introns. A major transcription initiation site was identified at the position 58 base pairs upstream of the AUG translation initiator codon on primer extension analysis with HeLa mRNA. The 5' flanking region lacks a typical TATA box but contains many putative binding sites for various transcription factors, such as Sp1, Oct1, NF1, AP1, E2F, and USF. Immediately downstream of the transcription start site, we found a VNTR of a 17-bp sequence rich in (G+C). Four different alleles with two to five copies of the tandem repeat were identified in 10 individuals examined, indicating a high degree of variation at the PTFgamma/SNAP43 locus. In addition, the PTFgamma/SNAP43 gene was mapped to human chromosome 14q22 by fluorescence in situ hybridization.

Base Sequence↗

PubMed1997

Insulin has a limited effect on the cell cycle progression in 3T3 L1 fibroblasts.

Insulin has pleiotropic effects on the regulation of cellular growth, differentiation, and metabolism. The biochemical events ultimately leading to cell proliferation after insulin treatment have been demonstrated in detail by numerous research groups. However, depending on cell types, it has been shown that insulin has various effects on cell proliferation. Therefore, we attempted to more critically evaluate the effect of insulin on cell proliferation in 3T3 L1 fibroblasts. In this study, we investigated insulin's effect on cell proliferation by using [3H]thymidine incorporation, flow cytometry, and cell counting. In 3T3 L1 fibroblasts studied in 0.5% serum, insulin induced a two-fold increase in [3H]thymidine incorporation over at 48 h, and the maximal rate of DNA synthesis was observed during 8-12 h incubation. The flow cytometric analysis also showed that insulin increased the cell population in the S phase. After insulin treatment for 48 h, cell numbers increased approximately 45% in comparison with 0.5% serum control. Cell division was found to occur only once in 60 h after staining 3T3 L1 fibroblasts with carboxyfluorescein diacetate succinimidyl ester (CFSE). Taken together, this data indicates that insulin stimulated the transit from the G0/G1 to S phase, progressed the cell cycle through the G2/M phase, and increased the cell number. However, under our experimental conditions, cells divided only once in 60 h in the presence of insulin.

3T3 Cells↗

PubMed1996

Isolation of guaianoid pigments from the gorgonian Calicogorgia granulosa.

Three pigments of the guaiazulene class have been isolated from the gorgonian Calicogorgia granulosa. Structures of these compounds have been determined as guaiazulene, 2,2'- diguaiazulenylmethane, and a new compound, 2,2'-biguaiazulenyl, by combined chemical and spectroscopic methods. 2,2'-Biguaiazulenyl exhibited moderate antimicrobial activity.

Animals↗

PubMed1996

Cloning of two proximal sequence element-binding transcription factor subunits (gamma and delta) that are required for transcription of small nuclear RNA genes by RNA polymerases II and III and interact with the TATA-binding protein.

The proximal sequence element (PSE)-binding transcription factor (PTF) specifically recognizes the PSEs of both RNA polymerase II- and RNA polymerase III-transcribed small nuclear RNA (snRNA) genes. We previously have shown that PTF purified from human HeLa cells is a multisubunit complex of four polypeptides designated PTF alpha, -beta, -gamma, and -delta. We now report the isolation and expression of cDNAs encoding PTF gamma and PTF delta, as well as functional studies with cognate antibodies that recognize the native PTF complex in HeLa extracts. Immunoprecipitation studies confirm that the four PTF subunits originally found to copurify during conventional chromatography indeed form a tightly associated complex; they further show that the PTF so defined, including the gamma and delta subunits specifically, is essential for transcription of both class II and class III snRNA genes. Immunoprecipitation assays also show a weak substoichiometric association of the TATA-binding protein (TBP) with PTF, consistent with the previous report of a PTF-related complex (SNAPc) containing substoichiometric levels of TBP and a component (SNAPc43) identical in sequence to the PTF gamma reported here. Glutathione S-transferase pulldown assays further indicate relatively strong direct interactions of both recombinant PTF gamma and PTF delta with TBP, consistent either with the natural association of TBP with PTF in a semistable TBP-TBP-associated factor complex or with possible functional interactions between PSE-bound PTF and TATA-bound TBP during promoter activation. In addition, we show that in extracts depleted of TBP and TBP-associated factors, transcription from the U1 promoter is restored by recombinant TBP but not by TFIID or TFIIIB, indicating that transcription of class II snRNA genes requires a TBP complex different from the one used for mRNA-encoding genes.

Amino Acid Sequence↗

PubMed1996

Cloning and characterization of the beta subunit of human proximal sequence element-binding transcription factor and its involvement in transcription of small nuclear RNA genes by RNA polymerases II and III.

The proximal sequence element (PSE)-binding transcription factor (PTF), which binds the PSE of both RNA polymerase II- and RNA polymerase III-transcribed mammalian small nuclear RNA (snRNA) genes, is essential for their transcription. We previously reported the purification of human PTF, a complex of four subunits, and the molecular cloning and characterization of PTF gamma and delta subunits. Here we describe the isolation and expression of a cDNA encoding PTF beta, as well as functional studies using anti-PTF beta antibodies. Native PTF beta, in either protein fractions or a PTF-Oct-1-DNA complex, can be recognized by polyclonal antibodies raised against recombinant PTF beta. Immunodepletion studies show that PTF beta is required for transcription of both classes of snRNA genes in vitro. In addition, immunoprecipitation analyses demonstrate that substantial and similar molar amounts of TATA-binding protein (TBP) and TFIIIB90 can weakly associate with PTF at low salt conditions, but this association is dramatically reduced at high salt concentrations. Along with our previous demonstration of both physical interactions between PTF gamma/PTF delta and TBP and the involvement of TFIIIB90 in the transcription of class III snRNA genes, these results are consistent with the notion that a TBP-containing complex related to TFIIIB is required for the transcription of class III snRNA genes, and acts through weak interaction with the four-subunit PTF.

Amino Acid Sequence↗

PubMed1996

The major histocompatibility complex class II Ea promoter requires TFIID binding to an initiator sequence.

The major histocompatibility complex (MHC) class II Ea promoter is dependent on the presence of conserved upstream X and Y boxes and of initiator (Inr) sequences. In vitro transcription analysis of the Inr region with linker-scanning mutants pinpoints a functionally essential element that shows homology to the terminal deoxynucleotidyltransferase (TdT) Inr; contrary to the TdT Inr and other Inrs identified so far, the key sequence, between positions +5 and +12, is located within a transcribed area. Swapping the TdT sequence into the corresponding Ea position leads to a fivefold increase in transcription rate, without altering start site selection. Inr-binding proteins LBP-1/CP2 and TIP--a TdT Inr-binding protein unrelated to YY1--recognize the Ea Inr; they interact with overlapping yet distinct sequences around the Cap site, but their binding does not coincide with Ea Inr activity. A good correlation is, rather, found with binding of immunopurified holo-TFIID to this element. TFIID interacts both with Ea TATA-like and Inr sequences, but only the latter is functionally relevant. Unlike TBP, TFIID binds in the absence of TFIIA, indicating a stabilizing role for TBP-associated factors in Ea promoter recognition. Sequence comparison with other mouse and human MHC class II promoters suggests a common mechanism of start site(s) selection for the MHC class II gene family.

Animals↗

PubMed1995

A novel LBP-1-mediated restriction of HIV-1 transcription at the level of elongation in vitro.

The cellular factor, LBP-1, can repress HIV-1 transcription by preventing the binding of TFIID to the promoter. Here we have analyzed the effect of recombinant LBP-1 on HIV-1 transcription in vitro by using a "pulse-chase" assay. LBP-1 had no effect on initiation from a preformed preinitiation complex and elongation to position +13 ("pulse"). However, addition of LBP-1 after RNA polymerase was stalled at +13 strongly inhibited further elongation ("chase") by reducing RNA polymerase processivity. Severe mutations of the high affinity LBP-1 binding sites between -4 and +21 did not relieve the LBP-1-dependent block. However, LBP-1 could bind independently to upstream low affinity sites (-80 to -4), suggesting that these sites mediate the effect of LBP-1 on elongation. These results demonstrate a novel function of LBP-1, restricting HIV-1 transcription at the level of elongation. In addition, Tat was found to suppress the antiprocessivity effect of LBP-1 on HIV-1 transcription in nuclear extracts. These findings strongly suggest that LBP-1 may provide a natural mechanism for restricting the elongation of HIV-1 transcripts and that this may be a target for the action of Tat in enhancing transcription.

Base Sequence↗

PubMed1995

Proximal sequence element-binding transcription factor (PTF) is a multisubunit complex required for transcription of both RNA polymerase II- and RNA polymerase III-dependent small nuclear RNA genes.

The proximal sequence element (PSE), found in both RNA polymerase II (Pol II)- and RNA Pol III-transcribed small nuclear RNA (snRNA) genes, is specifically bound by the PSE-binding transcription factor (PTF). We have purified PTF to near homogeneity from HeLa cell extracts by using a combination of conventional and affinity chromatographic methods. Purified PTF is composed of four polypeptides with apparent molecular masses of 180, 55, 45, and 44 kDa. A combination of preparative electrophoretic mobility shift and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses has conclusively identified these four polypeptides as subunits of human PTF, while UV cross-linking experiments demonstrate that the largest subunit of PTF is in close contact with the PSE. The purified PTF activates transcription from promoters of both Pol II- and Pol III-transcribed snRNA genes in a PSE-dependent manner. In addition, we have investigated factor requirements in transcription of Pol III-dependent snRNA genes. We show that in extracts that have been depleted of TATA-binding protein (TBP) and associated factors, recombinant TBP restores transcription from U6 and 7SK promoters but not from the VAI promoter, whereas the highly purified TBP-TBP-associated factor complex TFIIIB restores transcription from the VAI but not the U6 or 7SK promoter. Furthermore, by complementation of heat-treated extracts lacking TFIIIC activity, we show that TFIIIC1 is required for transcription of both the 7SK and VAI genes, whereas TFIIIC2 is required only for transcription of the VAI gene. From these observations, we conclude (i) that PTF and TFIIIC2 function as gene-specific as gene-specific factors for PSE-and B-box-containing Pol III genes, respectively, (ii) that the form of TBP used by class III genes with upstream promoter elements differs from the from used by class III genes with internal promoters, and (iii) that TFIIIC1 is required for both internal and external Pol III promoters.

Base Sequence↗

PubMed1994

Characterization of a family of related cellular transcription factors which can modulate human immunodeficiency virus type 1 transcription in vitro.

LBP-1 is a cellular protein which binds strongly to sequences around the human immunodeficiency virus type 1 (HIV-1) initiation site and weakly over the TATA box. We have previously shown that LBP-1 represses HIV-1 transcription by inhibiting the binding of TFIID to the TATA box. Four similar but distinct cDNAs encoding LBP-1 (LBP-1a, -b, -c, and -d) have been isolated. These are products of two related genes, and each gene encodes two alternatively spliced products. Comparison of the amino acid sequence of LBP-1 with entries in the available protein data bases revealed the identity of LBP-1c to alpha-CP2, an alpha-globin transcription factor. These proteins are also homologous to Drosophila melanogaster Elf-1/NTF-1, an essential transcriptional activator that functions during Drosophila embryogenesis. Three of the recombinant LBP-1 isoforms show DNA binding specificity identical to that of native LBP-1 and bind DNA as a multimer. In addition, antisera raised against recombinant LBP-1 recognize native LBP-1 from HeLa nuclear extract. Functional analyses in a cell-free transcription system demonstrate that recombinant LBP-1 specifically represses transcription from a wild-type HIV-1 template but not from an LBP-1 mutant template. Moreover, LBP-1 can function as an activator both in vivo and in vitro, depending on the promoter context. Interestingly, one isoform of LBP-1 which is missing the region of the Elf-1/NTF-1 homology is unable to bind DNA itself and, presumably through heteromer formation, inhibits binding of the other forms of LBP-1, suggesting that it may function as a dominant negative regulator.

Amino Acid Sequence↗

PubMed1993

Hepatic fatty acid-binding protein mRNA is regulated by growth hormone.

Hepatic fatty acid-binding protein (FABP) is one of several abundant proteins which may participate in fatty acid uptake and utilization. Using differential hybridization to screen for growth hormone-responsive gene products, a complementary deoxyribonucleic acid (cDNA) was isolated which proved to be a hepatic FABP cDNA fragment. Hypophysectomy caused a 60% reduction in hepatic FABP messenger ribonucleic acid (mRNA) levels in rat liver, and growth hormone administration to hypophysectomized rats resulted in restoration of the expression of hepatic FABP mRNA. Other pituitary hormones did not alter these changes in expression. The response to growth hormone occurred within 4 hours of administration. During development, expression of hepatic FABP mRNA in rat liver was low in late fetal life, with increases to 40% of adult values by day 2 of life. Significant increases to adult levels did not occur until after day 25, when weaning is essentially completed. Alteration of hepatic FABP mRNA expression by growth hormone in rat liver may be important in the complex regulation of fatty acid uptake and metabolism.

Animals↗

PubMed1992

Oct-1 and Oct-2 potentiate functional interactions of a transcription factor with the proximal sequence element of small nuclear RNA genes.

The promoters of both RNA polymerase II- and RNA polymerase III-transcribed small nuclear RNA (snRNA) genes contain an essential and highly conserved proximal sequence element (PSE) approximately 55 bp upstream from the transcription start site. In addition, the upstream enhancers of all snRNA genes contain binding sites for octamer-binding transcription factors (Octs), and functional studies have indicated that the PSE and octamer elements work cooperatively. The present study has identified and characterized a novel transcription factor (designated PTF) which specifically binds to the PSE sequence of both RNA polymerase II- and RNA polymerase III-transcribed snRNA genes. PTF binding is markedly potentiated by Oct binding to an adjacent octamer site. This potentiation is effected by Oct-1, Oct-2, or the conserved POU domain of these factors. In agreement with these results and despite the independent binding of Octs to the promoter, PTF and Oct-1 enhance transcription from the 7SK promoter in an interdependent manner. Moreover, the POU domain of Oct-1 is sufficient for significant in vitro activity in the presence of PTF. These results suggest that essential activation domains reside in PTF and that the potentiation of PTF binding by Octs plays a key role in the function of octamer-containing snRNA gene enhancers.

Base Sequence↗